Changes in Hepatocyte NADH Fluorescence during Prolonged Hypoxia

Obi-Tabot, Eliot; Hanrahan, Lawrence M.; Cachecho, Riad; Beer, Eve R.; Hopkins, Susan R.; Chan, Jeffery C.K.; Shapiro, Jerrold M.; LaMorte, Wayne W.

doi:10.1006/jsre.1993.1187

Cited by 58 publications

(30 citation statements)

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“…22 Ischemia has been well described to cause a physical reduction in NAD͑P͒H and a corresponding decrease in autofluorescence in a variety of cells and tissues. 23,45,46 The correlation with NADH autofluorescence intensity is related 47 to the physical concentration of the NADH. NADH autofluorescence has been used to assess the onset of skin flap necrosis in rats and showed a significant decrease in autofluorescence with time and necrosis.…”

Section: Discussionmentioning

confidence: 99%

“…46 When hepatocytes were exposed to longer hypoxic conditions, the autofluorescence gradually decreased from a peak level below the control until irreversible damage and cell death due to ischemia had occurred. 46 These changes occurred within 60 to 90 min after hypoxia. A similar trend was seen in this study, with the au- tofluorescence of the ex vivo skin initially increasing after 24 to 48 h incubation at 4°C and room temperature ͑Fig.…”

Section: Discussionmentioning

confidence: 99%

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Analysis of the metabolic deterioration of ex vivo skin from ischemic necrosis through the imaging of intracellular NAD(P)H by multiphoton tomography and fluorescence lifetime imaging microscopy

et al. 2010

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Abstract. Ex vivo human skin has been used extensively for cosmeceutical and drug delivery studies, transplantable skin allografts, or skin flaps. However, it has a half-life of a few days due to ischemic necrosis. Traditional methods of assessing viability can be timeconsuming and provide limited metabolic information. Using multiphoton tomography and fluorescence lifetime imaging ͑MPT-FLIM͒ we assess ischemic necrosis of ex vivo skin by NAD͑P͒H autofluorescence intensity and fluorescence lifetime. Ex vivo skin is stored in the presence and absence of nutrient media ͑Dulbecco Modified Eagle Medium͒ at −20, 4, and 37°C and room temperature over a 7-day time course to establish different rates of metabolic deterioration. At higher temperatures we observe a decrease in NAD͑P͒H autofluorescence, higher image noise, and a significant increase in the average fluorescence lifetime ͑ m ͒ from ϳ1000 to 2000 ps. Additionally, significant distortions in NAD͑P͒H fluorescence lifetime histograms correspond to the reduction in autofluorescence. Skin kept at 4°C, with or without media, showed the least change. Our findings suggest that MPT-FLIM enables useful noninvasive optical biopsies to monitor the metabolic state and deterioration of human skin for research and clinical purposes.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Analysis of the metabolic deterioration of ex vivo skin from ischemic necrosis through the imaging of intracellular NAD(P)H by multiphoton tomography and fluorescence lifetime imaging microscopy

et al. 2010

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“…Islets were spotted in Matrigel in four-quadrant dishes and treated similarly to INS-1 cells, although carbonyl cyanide 4-trifluoromethoxyphenylhydrazone was not used because of differences in basal (3 mM glucose) fluorescence within islets of the same condition. This was likely in part due to size differences among islets in which anoxia can be observed in the core of larger islets and may contribute to artificially high baseline levels of NADH (36). Thus, the scale that was used to analyze islets was fluorescence at basal (3 mM) glucose (0%, minimum NAD(P)H) and NaCN (100%, maximum NAD(P)H).…”

Section: Ins-1 Cellmentioning

confidence: 99%

Chronic Exposure to Excess Nutrients Left-shifts the Concentration Dependence of Glucose-stimulated Insulin Secretion in Pancreatic β-Cells

Erion

Berdan

Burritt

et al. 2015

Journal of Biological Chemistry

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Background: Fasting insulin secretion is increased in obesity and type 2 diabetes. Results: Culture of ␤-cells in excess nutrients resulted in increased lipid stores and a left-shifted dose-response curve of glucose-stimulated insulin secretion due to an enhanced sensitivity of exocytosis to Ca 2ϩ . Conclusion: Intracellular lipid modulates the dose response of glucose-stimulated insulin secretion. Significance: A shift in ␤-cell glucose sensitivity may contribute to hyperinsulinemia.

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“…A recent review (3) summarizing these studies shows that changes in NADH fluorescence due to oxygen deprivation have been measured in single cells (4), in tissue slices (5), and organs in vitro (6,7) and in vivo (8,9). Nevertheless, only a few studies have reported findings related to changes in NADH fluorescence due to reduction of oxygen supply in skin, the largest organ of the human body.…”

Section: Introductionmentioning

confidence: 99%

In Vivo Multiphoton NADH Fluorescence Reveals Depth-Dependent Keratinocyte Metabolism in Human Skin

Balu

Mazhar

Hayakawa

et al. 2013

Optics in the Life Sciences

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We employ a clinical multiphoton microscope to monitor in vivo and noninvasively the changes in reduced nicotinamide adenine dinucleotide (NADH) fluorescence of human epidermal cells during arterial occlusion. We correlate these results with measurements of tissue oxy-and deoxyhemoglobin concentration during oxygen deprivation using spatial frequency domain imaging. During arterial occlusion, a decrease in oxyhemoglobin corresponds to an increase in NADH fluorescence in the basal epidermal cells, implying a reduction in basal cell oxidative phosphorylation. The ischemia-induced oxygen deprivation is associated with a strong increase in NADH fluorescence of keratinocytes in layers close to the stratum basale, whereas keratinocytes from epidermal layers closer to the skin surface are not affected. Spatial frequency domain imaging optical property measurements, combined with a multilayer Monte Carlo-based radiative transport model of multiphoton microscopy signal collection in skin, establish that localized tissue optical property changes during occlusion do not impact the observed NADH signal increase. This outcome supports the hypothesis that the vascular contribution to the basal layer oxygen supply is significant and these cells engage in oxidative metabolism. Keratinocytes in the more superficial stratum granulosum are either supplied by atmospheric oxygen or are functionally anaerobic. Based on combined hemodynamic and two-photon excited fluorescence data, the oxygen consumption rate in the stratum basale is estimated to be~0.035 mmoles/10 6 cells/h.

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Changes in Hepatocyte NADH Fluorescence during Prolonged Hypoxia

Cited by 58 publications

References 0 publications

Analysis of the metabolic deterioration of ex vivo skin from ischemic necrosis through the imaging of intracellular NAD(P)H by multiphoton tomography and fluorescence lifetime imaging microscopy

Analysis of the metabolic deterioration of ex vivo skin from ischemic necrosis through the imaging of intracellular NAD(P)H by multiphoton tomography and fluorescence lifetime imaging microscopy

Chronic Exposure to Excess Nutrients Left-shifts the Concentration Dependence of Glucose-stimulated Insulin Secretion in Pancreatic β-Cells

In Vivo Multiphoton NADH Fluorescence Reveals Depth-Dependent Keratinocyte Metabolism in Human Skin

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